ISSN Journal of Pharmacognosy and Phytochemistry Psi - PDF document

ISSN 2278- 4136www.phytojournal.comJournal of Pharmacognosy and PhytochemistryVol.2012                                                 www.phytojournal.com Vikrant Arya, Narender ThakurCollege of Ayurvedic Pharmaceutical Sciences, Jogindernagar, Mandi H.P., India. [E-mail: arya.vikrant30@gmail.com] Vinayaka College of Pharmacy, Village Bahuguna P.O. Garsa Distt. Kullu, H.P., India Amritphale (Psidium guajava L.), is a small tree in Myrtaceae family, traditionally used in treatment of several diseases (inflammation, diabetes, hypertension, wounds, pain and fever). The present study was carried out to investigate the phytochemical profile of leaves of Psidium guajava L. The leaves powder was successively extracted flavonoids, tannins triterpenoids, saponins, sterols, alkaloids and carbohydrates. The result of the study could be useful for description and foundation of monograph of the plant. Flavonoids, Psidium, Phytochemical 1. Introduction Vikrant Arya*, Narender Thakur, C.P. KashyapVol.No.2012                                                 www.phytojournal.com                                                 Pagelayer of somewhat granular flesh, 1/8 to 1/2 inch thick, white, yellowish, light- or dark-pink, or near-red, juicy, acid, subacid, or sweet and flavorful. The central pulp, slightly darker in tone, is juicy and normally filled with very hard, yellowish seeds, 1/8 inch long, though some rare types have soft, chewable seeds. Actual seed counts have ranged from 112 to 535 but some guavas are seedless or nearly so. When immature and until a very short time before ripening, the fruit is green, hard, gummy within and very astringent. Bark is quite smooth, pale pinkish brown usually tinged with chlorophyll l . Taxonomic Classification More recent ethnopharmacological studies show that Psidium guajava is used in many parts of the world for the treatmentof a number of diseases, e.g. as an anti-inflammatory, for diabetes, hypertension, caries, wounds, pain relief and reducingfever. Some of the countries with a long history of traditionalmedicinal use of guava include Mexico and other CentralAmerican countries including the Caribbean, Africa and [6]2. Materials and method: 2.1 Collection of Plant Materials and Leaves: The leaves were collected from fields of Kangra, Himachal Pradesh, India. The collected leaves were shade dried for 35 days and finally pulverized in to coarse powder. It was stored in a well closed container free from environmental climatic changes till usage. 2.2 Preparation of Extracts: Successive extraction of plant material (Fig.1) Fig 1: A Schematic Representation of Successive extraction 2.3 Preparation of Hydroalcoholic Extract Hydroalcoholic (8:2) extract was prepared using leaves of Psidium guajava. Dried leaves (250 gm) were ground. The ground sample was soaked in hydroalcoholic solvent (8:2 v/v) and left for 24 h. The mixture was filtered and the filtrate concentrated by evaporation at 40 C. Then extract was dried and weighed. 2.4 Preliminary Phytochemical Screening The preliminary phytochemical tests were performed for testing different chemical groups present in extracts ts . Extract (100 mg) was treated with few drops of Dragendorff’s reagent [Potassium bismuth iodide solution]. Formation of orange brown precipitate indicated the presence of alkaloids. To 100 mg of extract small quantity of Wagner's reagent [Solution of iodine in potassium iodide] was added. Presence of reddish brown precipitate if alkaloids are present. To 100 mg of extract small quantity of Hager's Vikrant Arya*, Narender Thakur, C.P. KashyapVol.No.2012                                                 www.phytojournal.com                                                 Pagereagent [saturated solution of Picric acid] was added. Formation of yellow precipitate indicated the presence of alkaloids.Powdered drug extract on shaking vigorously with water results into persistent foam.200 mg extract was boiled with 3 mL of dil. H in a test tube for 5 min and filtered while hot. Cool and added the equal volume of C and CHCl, shake well and separated the organic solvent and added the NHThe ammonical layer turned pink or red. Alcoholic extract was treated with 1 mL pyridine and 1 mL of sodium nitroprusside. Pink to red colour appears. Extract (2 mL) was treated with 0.4 mL of glacial acetic acid containing a trace amount of FeCl and 0.5 mL of concentrated was also added by the side of the test tube. Persistent blue color appeared in the acetic acid layer if cardiac glycosides were present.To 5 mL of extract few drops of 5% FeClwas added. Presence of deep blue black colour indicated the presence of tannins. When 5 mL of extract was treated with few drops of 5% lead acetate solution, white precipitates appeared.To 5 mL of extract 5 mL of 95% ethanol was added along with dilute HCl from sides of test tube. Few fragments (0.5 g) of magnesium turnings were also added. Presence of slight pink colour indicated the presence of flavonoids. To 5 mL of extract few drops of NaOH solution was added. Formation of an intense yellow color, which turns to colorless on addition of few drops of dil. indicated the presence of flavonoids.little extract was taken with 2 mL of water and 0.5 mL of concentrated HNO was added to it. Yellow colour is obtained if proteins are present. To 5 mL of extract 4% NaOH was added along with few drops of 5% CuSO solution. Violet or pink colour appeared indicated the presence of proteins. Extract (5 mL) was treated with 5 mL CHCl with few drops of conc. H, shake well and allowed to stand for some time. Formation of yellow colored lower layer indicated the presence of triterpenoids. Extract (5 mL) was treated with few drops of acetic anhydride, boiled and cooled, conc. Hwas added from the sides of the test tube showed a brown ring at the junction of two layers and the upper layer turns green which showed the presence of Steroids and formation of deep red color indicated the presence of triterpenoids. In a test tube containing 5 mL of extract, few drops of freshly prepared 10% alcoholic solution of - naphthol was added and shaken/stirred for few min. Then 5 mL of conc. was added from sides of the test tube. Violet ring was formed at the junction of two liquids, indicated the presence of carbohydrates. Small quantity of extract was pressed the between two filter papers, the stain on I filter paper indicated the presence of fixed oils. The extract was evaporated to get 10 mL of extract. To the extract 25 mL of 10% NaOH was added, then it was boiled in water bath for 30 min. The extract was cooled and excess of sodium sulphate was added. Soap was formed at the top and filtered. To the filtrate H was added which was evaporated. The extract was dissolved in ethanol and few drops of CuSO and NaOH was added. Clear blue solution indicated the presence of fats. Observations: Successive and Hydroalcoholic Extraction of Powder of Psidium guajava Linn. Leaf Type of extract Amount of extract (gm) Yield (% ) Appearance Petroleum ether 2.850 1.54 Oily yellowish Chloroform 10.175 4.47 Greenish Ethanolic 18.013 7.61 Brownish black solid mass Aqueous 17.650 7.46 Brown Hydroalcoholic 20.425 8.57 Brown Vikrant Arya*, Narender Thakur, C.P. KashyapVol.2012                                                 www.phytojournal.com                                                 Page Table 2: Phytochemical Screening of Various Extracts of Psidium guajava (+) Positive Test, (-) Negative test 3. ConclusionPhytochemical screening of petroleum ether, chloroform, ethanol, aqueous and hydroalcoholic extracts revealed the presence flavonoids, tannins triterpenoids, saponins, sterols, alkaloids and carbohydrates by positive reaction with the respective test reagent. Phytochemical screening showed that maximum presence of phytoconstituents in ethanolic and hydroalcoholic extracts. 4. Acknowledgement:The authors are grateful to Mr. N.S Thakur, Chairman Vinyaka College of Pharmacy and Dr. C.P. Kashyap, Principal, College of Ayurvedic Pharmaceutical Sciences Jogindernagar, Mandi, Vikrant Arya*, Narender Thakur, C.P. KashyapVol.No.2012                                                 www.phytojournal.com                                                 PageH.P. for providing facilities to carrying out research on Psidium leaves. 5. 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